Methods for the early diagnosis of ovarian cancer

Abstract

The present invention discloses the protease hepsin is specifically over-expressed in ovarian and other malignancies. A number of hepsin peptides can induce immune responses to hepsin, thereby demonstrating the potential of these peptides in monitoring and the development of immunotherapies for ovarian and other malignancies. There is also provided a hepsin protein variant that is useful as a marker for ovarian cancer cells, prostate cancer cells or kidney cancer cells.

1 . An isolated DNA encoding a hepsin protein variant, said variant protein comprising an amino acid sequence shown in SEQ ID NO. 195 or a peptide variant thereof. 
     
     
         2 . The isolated DNA of  claim 1 , wherein the peptide variant amino acid sequence is shown in SEQ ID NO: 196. 
     
     
         3 . An isolated and purified hepsin protein variant with an amino acid sequence shown in SEQ ID NO. 195 or a hepsin protein variant peptide thereof with an amino acid sequence shown in SEQ ID NO: 196. 
     
     
         4 . An immunogenic composition, comprising the hepsin protein variant or peptide thereof of  claim 3  and an appropriate adjuvant. 
     
     
         5 . A method of vaccinating an individual against hepsin protein variant, comprising:
 inoculating an individual with the hepsin protein variant or peptide thereof of  claim 3 , wherein said hepsin protein variant or peptide elicits an immune response in the individual, thereby vaccinating the individual against hepsin protein variant.   
     
     
         6 . The method of  claim 5 , wherein said hepsin protein variant or peptide thereof comprises a dendritic cell or is expressed from an expression vector. 
     
     
         7 . The method of  claim 5 , wherein the individual has or is at risk of getting ovarian cancer, lung cancer, prostate cancer, or colon cancer. 
     
     
         8 . A method of immunotherapy targeted toward hepsin protein variant in an individual, comprising:
 a) isolating dendritic cells from the individual;   b) expressing a hepsin protein variant or peptide variant thereof of  claim 3  in the dendritic cells;   c) exposing immune cells comprising T cells isolated from the individual to the dendritic cells, wherein said dendritic cells would generate hepsin variant-specific T cells from the immune cells; and   d) transferring the immune cells back to the individual, wherein said immune cells would activate hepsin variant-specific immune responses in the individual, thereby generating immunotherapy targeted toward hepsin protein variant.   
     
     
         9 . The method of  claim 8 , wherein the individual has, is suspected of having or is at risk of getting ovarian cancer or prostate cancer. 
     
     
         10 . The method of  claim 8 , wherein expression of hepsin protein variant in the dendritic cells is obtained by transfection, transduction and loading the dendritic cells with a hepsin protein variant or fragment thereof. 
     
     
         11 . A method of monitoring the efficacy of vaccinating an individual with hepsin protein variant or fragment thereof, comprising:
 vaccinating the individual with the hepsin protein variant or peptide variant thereof of  claim 3 ;   isolating T cells from the individual; and   measuring immune responses induced by the hepsin variant or fragment thereof, wherein an increased level of immune responses compared to those exhibited by cells from normal individual indicates that the individual has been vaccinated by said hepsin protein variant or fragment thereof.   
     
     
         12 . The method of  claim 11 , wherein the immune response is T cell proliferation induced by the hepsin protein variant or peptide variant thereof, frequency of cytokine-secreting T cells specific to the hepsin protein variant or peptide variant thereof and frequency of T cells expressing T cell receptor specific to the hepsin protein variant or peptide variant thereof. 
     
     
         13 . A method of inhibiting hepsin protein variant in a cell, comprising
 introducing into the cell an antibody specific for a hepsin protein variant of  claim 3 , wherein binding of the antibody to the hepsin protein variant or peptide variant thereof inhibits hepsin protein variant in the cell.   
     
     
         14 . The method of claim, further comprising a therapeutic moiety. 
     
     
         15 . The method of  claim 13 , wherein the therapeutic moiety is a radioisotope, a toxin, a chemotherapeutic agent, an immune stimulant, or a cytotoxic agent. 
     
     
         16 . The method of  claim 13 , wherein the individual has ovarian cancer or prostate cancer. 
     
     
         17 . A method of producing immune-activated cells directed toward a hepsin protein variant, comprising:
 exposing dendritic cells to a hepsin protein variant with a sequence shown in SEQ ID NO: 195 or peptide variant therefrom, wherein the exposure activates the immune cells and produces immune-activated cells directed toward the hepsin protein variant.   
     
     
         18 . The method of  claim 17 , wherein the peptide variant amino acid sequence is shown in SEQ ID NO: 196 
     
     
         19 . The method of  claim 17 , wherein the dendritic cells are isolated from an individual prior to the exposure, wherein said activated dendritic cells are reintroduced into the individual subsequent to the exposure. 
     
     
         20 . The method of  claim 17 , wherein the individual has or is at risk of getting ovarian cancer, lung cancer, prostate cancer, or colon cancer.